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Cross-Linked Enzyme Aggregates of Phenylalanine Ammonia Lyase: Novel Biocatalysts for Synthesis of L-Phenylalanine

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Abstract

Cross-linked enzyme aggregates of phenylalanine ammonia lyase (PAL-CLEAs) from Rhodotorula glutinis were prepared. The effects of the type of aggregating agent, its concentration, and that of cross-linking agent were studied. PAL-CLEAs production was most effective using ammonium sulfate (40 % saturation), followed by cross-linking for 1 h with 0.2 % (v/v) glutaraldehyde. Moreover, the storage and operational stability of the resulting PAL-CLEAs were also investigated. Compared to the free enzyme, the PAL-CLEAs exhibited the expected increased stability of the enzyme against various deactivating conditions such as pH, temperature, denaturants, and organic solvents and showed higher storage stability than its soluble counterpart. Additionally, the reusability of PAL-CLEAs with respect to the biotransformation of l-phenylalanine was evaluated. PAL-CLEAs could be recycled at least for 12 consecutive batch reactions without dramatic activity loss, which should dramatically increase the commercial potential of PAL for synthesis of l-phenylalanine. To the best of our knowledge, this is the first report of immobilization of PAL as cross-linked enzyme aggregates.

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Acknowledgments

The project was partially supported by the National Natural Science Foundation of China (NSFC, Project No. 21072041), Open Funding Project of the National Key Laboratory of Biochemical Engineering (no. KF2010-12) and the Foundation (no. GYWSW02) of Tianjin Key Laboratory of Industrial Microbiology (Tianjin University of Science and Technology), People’s Republic of China, Natural Science Foundation of Hebei University of Science and Technology (NO. XL201149), and Foundation of Hebei University of Science and Technology for Distinguished Young Scientists.

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  1. Research Center for Fermentation Engineering of Hebei, College of Bioscience and Bioengineering, Hebei University of Science and Technology, 70 Yuhua East Road, Shijiazhang, 050018, People’s Republic of China

    Jian-Dong Cui, Si Zhang & Li-Mei Sun

  2. National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, HaiDian District, Beijing, 100080, People’s Republic of China

    Jian-Dong Cui

  3. Key Laboratory of Industry Microbiology, Tianjin University of Science and Technology, Ministry of Education, 29 Thirteenth Street, Tai Da Development Area, Tianjin, 300457, People’s Republic of China

    Jian-Dong Cui

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  1. Jian-Dong Cui
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  2. Si Zhang
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Correspondence to Jian-Dong Cui.

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Cui, JD., Zhang, S. & Sun, LM. Cross-Linked Enzyme Aggregates of Phenylalanine Ammonia Lyase: Novel Biocatalysts for Synthesis of L-Phenylalanine. Appl Biochem Biotechnol 167, 835–844 (2012). https://doi.org/10.1007/s12010-012-9738-0

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  • DOI: https://doi.org/10.1007/s12010-012-9738-0

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